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Potapenko, E.

Publications and source records attributed to Potapenko, E..

2 recordsLinked to original sources

The Role of Host Calcium Signaling in Toxoplasma gondii egress

Toxoplasma gondii, an obligate intracellular parasite, is capable of invading virtually any nucleated cell. Ca2+ signaling is universal and both T. gondii and its mammalian host cell will utilize Ca2+ signaling to stimulate diverse cellular functions. Egress of T. gondii from the host cell is an essential step for the infection cycle of T. gondii and a cytosolic Ca2+ increase initiates the Ca2+ signaling cascade that culminates in stimulation of motility and egress. In this work we demonstrate that intracellular T. gondii is capable of taking up Ca2+ from the host cytoplasm when this concentration is increased during host signaling events. Both intracellular and extracellular Ca2+ sources are important to reach a threshold of cytosolic Ca2+ needed for a successful egress. Two peaks of Ca2+ were observed in single parasites that egressed with the second peak resulting from Ca2+ influx. We patched infected host cells to allow a precise delivery of exact concentrations of Ca2+ for stimulating motility and egress. Using this approach, we found that low potassium concentration modulates but do not trigger host cell egress. This is the first study using whole-cell patches to study the role of ions such as K+ and Ca2+ in T. gondii egress.

microbiology

Pyruvate kinase controls signal strength in the insulin secretory pathway

Pancreatic {beta}-cells couple nutrient metabolism with appropriate insulin secretion. Here, we show that pyruvate kinase (PK), which converts ADP and phosphoenolpyruvate (PEP) into ATP and pyruvate, underlies {beta}-cell sensing of both glycolytic and mitochondrial fuels. PK present at the plasma membrane is sufficient to close KATP channels and initiate calcium influx. Small-molecule PK activators increase {beta}-cell oscillation frequency and potently amplify insulin secretion. By cyclically depriving mitochondria of ADP, PK restricts oxidative phosphorylation in favor of the mitochondrial PEP cycle with no net impact on glucose oxidation. Our findings support a compartmentalized model of {beta}-cell metabolism in which PK locally generates the ATP/ADP threshold required for insulin secretion, and identify a potential therapeutic route for diabetes based on PK activation that would not be predicted by the {beta}-cell consensus model. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=164 HEIGHT=200 SRC="FIGDIR/small/907790v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@1f6503forg.highwire.dtl.DTLVardef@e9393org.highwire.dtl.DTLVardef@fab08borg.highwire.dtl.DTLVardef@5c7c55_HPS_FORMAT_FIGEXP M_FIG C_FIG The consensus model for {beta}-cell glucose sensing supports a dominant role for OxPhos. This model doesnt fully explain the observed metabolic and electrophysiologic oscillations associated with glucose-stimulated insulin secretion. Lewandowski et al. challenge this model by mechanistically connecting the anaplerotic PEP cycle to the electrically silent triggering phase, and OxPhos to the electrically active secretory phase. Here, the allosteric recruitment of pyruvate kinase directs metabolic traffic between the two cycles and identifies potential therapeutic strategies for diabetes based on pharmacologic pyruvate kinase activation. HIGHLIGHTSO_LICompartmentalized pyruvate kinase (PK) activity underlies {beta}-cell fuel sensing C_LIO_LIMembrane-associated PK closes KATP channels and controls calcium influx C_LIO_LIBy lowering ADP, PK toggles mitochondria between OxPhos and PEP biosynthesis C_LIO_LIPharmacologic PK activation increases oscillatory frequency and amplifies secretion C_LI

cell biology